Dreisbach J H, Merkel J R
J Bacteriol. 1978 Aug;135(2):521-7. doi: 10.1128/jb.135.2.521-527.1978.
The inducible nature of an extracellular collagenase produced by a marine Vibrio (Vibrio B-30, ATCC 21250) was demonstrated by observing the increase in extracellular collagenase activity after the addition of collagen to cell cultures in the latter part of the exponential growth phase. When collagenase-hydrolyzed collagen was added, the lag time required before collagenase production was detected decreased significantly compared with cultures receiving collagen. Cells preinduced to synthesize collagenase did not produce the enzyme when collagen was removed from the culture medium. Incorporation of penicillin G had no effect on final collagenase activity levels in suspensions of Vibrio B-30 in complete medium supplemented with collagen. However, chloramphenicol and tetracycline inhibited collagenase production, indicating that de novo protein synthesis was necessary for the appearance of activity. Attempts to isolate the inducing substance(s) involved filtering hydrolyzed collagen through a series of ultrafiltration membranes. The lowest-molecular-weight fraction of collagen hydrolysate with inducing ability was between 1,000 and 10,000. Gel filtration of this fraction on Sephadex G-50 resulted in the appearance of three protein peaks, two of which were capable of inducing collagenase production. Results from amino acid composition and N-terminal amino acid analysis suggest that the inducing substance originates from the polar helical portion of the collagen molecule.
通过观察在指数生长期后期向细胞培养物中添加胶原蛋白后细胞外胶原酶活性的增加，证明了海洋弧菌（弧菌B - 30，ATCC 21250）产生的细胞外胶原酶的可诱导性质。当添加胶原酶水解的胶原蛋白时，与接受胶原蛋白的培养物相比，检测到胶原酶产生所需的延迟时间显著缩短。当从培养基中去除胶原蛋白时，预先诱导合成胶原酶的细胞不会产生该酶。在补充有胶原蛋白的完全培养基中，青霉素G的加入对弧菌B - 30悬浮液中的最终胶原酶活性水平没有影响。然而，氯霉素和四环素抑制胶原酶的产生，表明从头合成蛋白质对于活性的出现是必需的。尝试通过一系列超滤膜过滤水解胶原蛋白来分离相关的诱导物质。具有诱导能力的胶原蛋白水解产物的最低分子量部分在1000至10000之间。该部分在Sephadex G - 50上进行凝胶过滤后出现三个蛋白质峰，其中两个能够诱导胶原酶的产生。氨基酸组成和N端氨基酸分析结果表明，诱导物质源自胶原蛋白分子的极性螺旋部分。
